Protocol for Tri-culture of hiPSC-Derived Neurons, Astrocytes, and Microglia

This protocol establishes a tri-culture of hiPSC-derived neurons, astrocytes, and microglia for the study of cellular interactions during homeostasis, injury, and disease. This system allows for mechanistic studies that can identify the roles of individual cell types in disease and injury response in a physiologically relevant, all-human system. This protocol utilizes and modifies prior differentiations. Limitations include the prolonged maturation of human astrocytes and neurons and scalability. For complete details on the use and execution of this protocol, please refer to Ryan et al. (2020).